- Email: [email protected]
417 LOSS OF C-MYC DOES NOT PREVENT PROLIFERATION IN HEPATOCYTES
POSTERS levels and elevated bilirubin levels. These alterations are paralleled by impaired glucose tolerance in transgenic versus wild type mice. Measurement of liver-specific enzymes (ALT and AST) showed slightly elevated levels. Conclusions: These results show that Fox O3 and insulin signaling in the liver is critical in regulating glucose homeostasis and maintaining normal hepatic function. 415 HEPATIC STELLATE CELL AS A SOURCE FOR HEPATOCYTE DIFFERENTIATION BUT NO HEPATIC FIBROSIS DEVELOPMENT A. Gumerova, A. Shafigullina, A. Trondin, M. Titova, M. Kaligin, I. Gazizov, D. Andreeva, S. Abdulkhakov, A. Kiassov. Department of Normal Human Anatomy, Kazan State Medical University, Kazan, Russia E-mail: [email protected] Background and Aim: Wide spread of liver diseases and low efficacy of their treatment require to search new ways for solving this problem. So, the one of promising method can be identification and application of hepatic progenitor cell. Because hepatic stellate cells (HSC) produce important morphogenic cytokines (Hepatocyte Growth Factor, Stem Cell Factor, ets.) and express stem cell markers, the aim of our study is to check the possibility of hepatocyte differentiation of HSC. Materials and Methods: We studied capability of HSC to hepatocyte differentiation with immunohisto- and cytochemically methods: 1. during rat and human prenatal development (human liver were obtained after legal medical abortions); 2. during liver regeneration after partial hepatectomy or toxic damage with lead nitrate; 3. in pure culture and in culture with addition of growth factors provided primary differentiation of hepatocytes in embryogenesis (Fibroblast Growth Factor, Hepatocyte Growth Factor); 4. after transplantation of HSC transfected with Green Fluorescent Protein (GFP) into intact rats and rats after partial hepatectomy (PH). Results: We found that HSC expressed stem cell markers both during the first trimester of human gestation (Bcl-2) and regeneration in rats (C-kit). Moreover they were cytokeratin18+/cytokeratin19+ within the embryonic period and after formation of monolayer in culture too (21 day; 1–3 passages). In the latter case HSC express a-fetoprotein and g-gamma-glutamyl transpeptidase also. Growth factors in culture intensified and accelerated the appearance of cytokeratins, a-Fetoprotein and Hepatocyte Specific Antigen in HSC. So HSC could be become hepatocyte-like cells. At last we observed GFP+ hepatocytes in rat’s livers after transplantation of GFP-transfected HSC to them, and there were more numerous GFP+ hepatocytes in livers after PH. So, PH accelerates repopulation of liver by HSC. There weren’t GFP+ myofibroblasts or fibrosis in all these livers. Conclusions: HSC can differentiate into hepatocytes both in vivo and in vitro. Thus these cells have properties and potencies of hepatic progenitor cells and they can be used for elaboration of stem cell therapy methods in hepatology.
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416 CELLULAR IMMUNE RESPONSE FOLLOWING HUMAN FETAL LIVER-DERIVED STEM CELLS TRANSPLANTATION IN CASE OF DECOMPENSATED LIVER CIRRHOSIS M.A. Habeeb1 , A.A. Khan2 , G. Sivaram2 , A. Bardia2 , G. Srinivas3 , T. Avinashraj3 , G. Pande3 . 1 Department of Gastroenterology and Hepatology, 2 Department of Gastroenterology & Hepatology, Centre for Liver Research & Diagnostics, Deccan College of Medical Sciences, 3 Centre for Cellular and Molecular Biology, Hyderabad, India E-mail: [email protected] Liver transplantation is the only effective treatment for decompensated liver-cirrhosis. Several factors, such as nonavailability of donors, operative-risks, complications associated with rejection, usage of immunosuppressive agents, and high cost of treatment, make this strategy available to only a few people. Hepatic progenitor stem cell transplantation (HSCT) using human fetal liverderived stem cells have been shown to be a good alternative to manage end-stage liver diseases. In this retrospective study, we investigated safety and efficacy of HSCT by monitoring the T-cell, NK-cell and cytokines which play major role in cellular immune response and rejection of chronic decompensated liver cirrhosis patients. The study protocol was approved by Institutional Ethics Committee, Deccan College of Medical Sciences, Hyderabad, a total of 5 patients with decompensated liver cirrhosis were enrolled in the study. After giving human fetal liver-derivedEPCAM positive cell transplantation, T-cell (CD3, CD4 and CD8), NK-cells (CD16)by flowcytometry and cytokine-levels (IL2, TNFb, IFNa, INFb and IFNg) by ELISA were monitored four times within a month. Present study demonstrated that after HSCT patient showed marked clinical recovery and decline in the MELD score and there was no significant variation found in cell mediated response and cytokine levels between pre and post transplantation. Hence this preliminary study demonstrated human fetal liver-derived EPCAM positive stem cell transplantation is safety for end stage liver cirrhosis. 417 LOSS OF C-MYC DOES NOT PREVENT PROLIFERATION IN HEPATOCYTES K. Hanak, S. Marhenke, M. Manns, A. Vogel, A.G. Vogel. Gastroenterology, Hepatology & Endocrinology, Hannover Medical School, Hannover, Germany E-mail: [email protected] Background and Aims: Myc is involved in many cellular processes as cell cycle progression and cell growth but also differentiation and apoptosis. Deregulation of myc may lead to uncontrolled cell proliferation, genomic instability, independence of growth factors and tumour development, especially in HCC, where myc has been identified to play a critical role. Hereditary Tyrosinemia type 1 is a genetic defect in the tyrosine-decomposition. The toxic metabolite Fumarylacetoacetate (FAA) accumulates and leads to hepatitis and a following tumour development by 6 years of age in 36% of all patients. NTBC (2-(2-N-4-trifluoromethylbenzoyl)-1,3cyclohexanedione) blocks the tyrosine-catabolism upstream FAA and is an effective treatment to prevent the hepatitis. The Fahknockout mouse-model mimics the human disease: Fah−/− -mice develop a severe liver damage after NTBC-withdrawal and die within 8 weeks. Under a low-dose NTBC-treatment (2.5%) they develop a chronic hepatitis and HCCs within 9 months. Methods: To analyse the role of c-myc in chronic liver diseases, Fah−/− -mice were crossed with c-mycfl/fl mxCre+ -mice. C-myc was deleted by Polyinosinic-Polycytodylic Acid-injections in 4-week old Fah−/− c-mycfl/fl -mice. To analyse c-myc function in liver injury, three approaches were used: 1. Mice were taken 0% NTBC for 2 weeks (acute liver injury). 2. Mice were kept under 2.5% NTBC-treatment (chronic hepatitis) for 4 weeks. 3. Flares of liver injury were induced by NTBC-cycling (21 days 0% and 5 days 100% NTBC) to mimic liver
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POSTERS damage with subsequent liver regeneration. Blood and livers were collected and analysed. Results: Loss of c-myc did not modulate apoptosis and proliferation of hepatocytes during severe and moderate liver injury. Similarly to Fah−/− -mice, Fah−/− c-myc−/− -double-knockout mice displayed few BrdU/Ki67- and TUNEL-positive hepatocytes. Surprisingly, liver regeneration was significantly enhanced in c-myc-deficient mice following NTBC-cycling. Interestingly, more proliferating hepatocytes were observed in Fah−/− c-myc−/− -double-knockout mice in the regeneration-phase despite reduced liver damage in histological and biochemical analysis. Currently, the molecular mechanism of liver regeneration by c-myc is investigated. Conclusions: C-myc does not affect liver injury in Fah−/− -mice during severe and moderate liver-damage. Surprisingly, loss of c-myc ameliorates liver injury and enhances liver regeneration during repeated flares of liver injury. Long-term studies will reveal c-myc’s contribution to hepatocarcinogenesis in the Fah-model. 418 SEPSIS INDUCED DAMAGE IN THE LIVER IS MODULATED BY PEROXIREDOXINS N. Huber1 , T. Annecke2 , A. Hillenbrand1 , D. Henne-Bruns1 , U. Knippschild1 , J. Tschop ¨ 2 . 1 General-, Visceral- and Transplantation Surgery, University of Ulm, Ulm, 2 Anesthesiology, University of Munich, Munich, Germany E-mail: [email protected] Background and Aims: Severe sepsis is characterized by hyperinflammation, oxidative damage, hypercoagulation, tissue hypoperfusion, hypoxia, and immune suppression, leading to multiorgan dysfunction. During sepsis, liver dysfunction is common and contributes to high mortality. Although the precise mechanisms by which the liver is affected are unclear, evidence is increasing that failure of mitochondria to effectively couple oxygen consumption with energy production contribute to the pathology of sepsis. An imbalance between formation and clearance of reactive oxygen species (ROS) results in oxidative stress and significantly contributes to dysfunction of cells during sepsis. In addition, ROS are essential to various functions of cells. However, moderate levels of antioxidant defenses are required to protect against the harmful effects of excessive ROS production. The aim of this study was to evaluate the levels of antioxidants, especially those of peroxiredoxin (Pdrx) isoforms 3 and 6, during induced sepsis in a mouse model. Material and Methods: For all experiments 8–12 weeks old male C57BL/6 mice either on a normal or high fat diet were used. Mice were randomized for the groups and all surgical procedures were performed in a matched experimental setting. Sepsis was induced by cecal ligation and puncture method. Animals were sacrificed 6 h, 12 h, and 24 h after induction of sepsis. Changes in the expression of various proteins were analyzed by Western Blot analyses and immunhistochemistry. Additionally, Kaplan-Meieranalysis were performed. Results: Already 6 h upon induction of sepsis changes in the cytoplasm of the liver could be detected. The expression levels of the relatively new group of antioxidants, the peroxiredoxins (Prdx 3 and 6) in the liver were detected by Western blot analyses and immunohistochemistry. Furthermore, we show that the influence of nutrition modulate the levels of Prdx proteins in the liver thereby influencing the severity of sepsis and survival. In addition, our results show that changes in the expression of Prdx isoforms modulate death signalling. Conclusions: Here we show for the first time that changes of Pdrx levels in the liver depend on the severity of sepsis. These results might contribute to the development of new therapeutical strategies in the treatment of severe sepsis.
419 RESULTS OF AUTOLOGOUS BONE MARROW CELL INFUSION IN PATIENTS WITH ADVANCED LIVER CIRRHOSIS J.K. Kim1,2 , H.J. Chung1 , D.H. Jung1 , D.Y. Kim1,2 , S.H. Ahn1,2 , C.Y. Chon1 , K.-H. Han1,2 , K.S. Lee1,2 . 1 Dept. of Internal Medicine, Yonsei University College of Medicine, 2 Liver Cirrhosis Clinical Research Center, Seoul, Republic of Korea E-mail: [email protected] Background and Aim: Liver cirrhosis (LC) is the end stage of chronic liver disease and is very difficult to treat. Although human clinical trials of bone marrow cell (BMC) infusion for cirrhotic patients showed positive results, the long-term follow up data after the procedure is limited. The aim of this study is to analyze the clinical results of phase II study after autologous BMC infusion (ABMI) in patients with advanced LC. Patients and Methods: Patients aged between 18 and 75 with a clinical diagnosis of advanced LC (Child-Pugh class B) and no viable hepatocellular carcinoma on magnetic resonance imaging (MRI) underwent ABMI. Autologous BMCs were harvested from the ilium under general anesthesia, and infused into peripheral vein after RBC depletion and mononuclear cell concentration. Serologic tests and questionnaires about performance were repeated during the 12-month period after the ABMI. Data after liver transplantation were not included. Results: Twenty patients were screened and 19 patients were enrolled. Mean age was 52 year-old. Patients (M:F = 9:10) were followed for more than 12 months after ABMI. Eighteen patients had B-viral liver cirrhosis and one had alcoholic liver cirrhosis. Serum albumin level was improved from 2.9 (2.3–3.6) g/dL before ABMI to 3.4 (2.6–4.5) g/dL at 12th month after ABMI. Prothrombin time showed improving tendency from 70 (60.0–84.0) % before ABMI to 77 (69.0–100) % at 12th month. Hemoglobin was increased from 11.6 (9.5–12.5) g/dL before ABMI to 13.25 (11.4–16.8) g/dL at 12th month. Child-Pugh score was decreased from 7.6 (7–12) before ABMI to 6.8 (5–10) at 12th month after ABMI. Performance status score improved from 78.9 (70–80) before ABMI to 90.8 (70–100) at 12th month. Conclusions: ABMI for advanced liver cirrhosis improved hepatic function and ABMI can be a bridge therapy waiting liver transplantation. 420 NON-PARENCHYMAL CELLS REGULATE PROLIFERATION AND DIFFERENTIATION OF HUMAN LIVER PROGENITOR CELLS J.W.C. Kung, I.S. Currie, P.W. Hadoke, S.J. Forbes, J.A. Ross. University of Edinburgh, Edinburgh, UK E-mail: [email protected] Background: Human foetal liver progenitor cells (LPCs) have promise for liver cell therapy, however, culture of LPCs alone result in inadequate functionality and phenotypic stability. The conditions required for LPC proliferation and differentiation are currently unknown. We hypothesised that non-parenchymal cells (NPCs) regulate proliferation and differentiation of LPCs. Methods: EpCAM+ LPCs were isolated by immunoselection from second trimester human livers. NPCs were isolated through EpCAM negative selection. LPCs, NPCs and mixed LPC/NPC cultures were grown in Williams’ Medium E (WME) and NPC-conditioned medium (NPC-CM) for 13 days and their hepatocytic functions were compared. Results: EpCAM+ cells had hepatoblast phenotype (AFP/Albumin/ CK19 positive). EpCAM− NPCs consisted of tube forming endothelial cells and mesenchymal cells. Mixed LPC/NPC cultures had significantly greater synthetic competence than LPCs alone (twoway ANOVA, p < 0.01). We confirmed this was mediated through soluble factor(s) by performing media transfers. LPCs grown in NPC-CM had greater hepatocytic maturity by secreting significantly
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416 CELLULAR IMMUNE RESPONSE FOLLOWING HUMAN FETAL LIVER-DERIVED STEM CELLS TRANSPLANTATION IN CASE OF DECOMPENSATED LIVER CIRRHOSIS M.A. Habeeb1 , A.A. Khan2 , G. Sivaram2 , A. Bardia2 , G. Srinivas3 , T. Avinashraj3 , G. Pande3 . 1 Department of Gastroenterology and Hepatology, 2 Department of Gastroenterology & Hepatology, Centre for Liver Research & Diagnostics, Deccan College of Medical Sciences, 3 Centre for Cellular and Molecular Biology, Hyderabad, India E-mail: [email protected] Liver transplantation is the only effective treatment for decompensated liver-cirrhosis. Several factors, such as nonavailability of donors, operative-risks, complications associated with rejection, usage of immunosuppressive agents, and high cost of treatment, make this strategy available to only a few people. Hepatic progenitor stem cell transplantation (HSCT) using human fetal liverderived stem cells have been shown to be a good alternative to manage end-stage liver diseases. In this retrospective study, we investigated safety and efficacy of HSCT by monitoring the T-cell, NK-cell and cytokines which play major role in cellular immune response and rejection of chronic decompensated liver cirrhosis patients. The study protocol was approved by Institutional Ethics Committee, Deccan College of Medical Sciences, Hyderabad, a total of 5 patients with decompensated liver cirrhosis were enrolled in the study. After giving human fetal liver-derivedEPCAM positive cell transplantation, T-cell (CD3, CD4 and CD8), NK-cells (CD16)by flowcytometry and cytokine-levels (IL2, TNFb, IFNa, INFb and IFNg) by ELISA were monitored four times within a month. Present study demonstrated that after HSCT patient showed marked clinical recovery and decline in the MELD score and there was no significant variation found in cell mediated response and cytokine levels between pre and post transplantation. Hence this preliminary study demonstrated human fetal liver-derived EPCAM positive stem cell transplantation is safety for end stage liver cirrhosis. 417 LOSS OF C-MYC DOES NOT PREVENT PROLIFERATION IN HEPATOCYTES K. Hanak, S. Marhenke, M. Manns, A. Vogel, A.G. Vogel. Gastroenterology, Hepatology & Endocrinology, Hannover Medical School, Hannover, Germany E-mail: [email protected] Background and Aims: Myc is involved in many cellular processes as cell cycle progression and cell growth but also differentiation and apoptosis. Deregulation of myc may lead to uncontrolled cell proliferation, genomic instability, independence of growth factors and tumour development, especially in HCC, where myc has been identified to play a critical role. Hereditary Tyrosinemia type 1 is a genetic defect in the tyrosine-decomposition. The toxic metabolite Fumarylacetoacetate (FAA) accumulates and leads to hepatitis and a following tumour development by 6 years of age in 36% of all patients. NTBC (2-(2-N-4-trifluoromethylbenzoyl)-1,3cyclohexanedione) blocks the tyrosine-catabolism upstream FAA and is an effective treatment to prevent the hepatitis. The Fahknockout mouse-model mimics the human disease: Fah−/− -mice develop a severe liver damage after NTBC-withdrawal and die within 8 weeks. Under a low-dose NTBC-treatment (2.5%) they develop a chronic hepatitis and HCCs within 9 months. Methods: To analyse the role of c-myc in chronic liver diseases, Fah−/− -mice were crossed with c-mycfl/fl mxCre+ -mice. C-myc was deleted by Polyinosinic-Polycytodylic Acid-injections in 4-week old Fah−/− c-mycfl/fl -mice. To analyse c-myc function in liver injury, three approaches were used: 1. Mice were taken 0% NTBC for 2 weeks (acute liver injury). 2. Mice were kept under 2.5% NTBC-treatment (chronic hepatitis) for 4 weeks. 3. Flares of liver injury were induced by NTBC-cycling (21 days 0% and 5 days 100% NTBC) to mimic liver
Journal of Hepatology 2012 vol. 56 | S71–S224
POSTERS damage with subsequent liver regeneration. Blood and livers were collected and analysed. Results: Loss of c-myc did not modulate apoptosis and proliferation of hepatocytes during severe and moderate liver injury. Similarly to Fah−/− -mice, Fah−/− c-myc−/− -double-knockout mice displayed few BrdU/Ki67- and TUNEL-positive hepatocytes. Surprisingly, liver regeneration was significantly enhanced in c-myc-deficient mice following NTBC-cycling. Interestingly, more proliferating hepatocytes were observed in Fah−/− c-myc−/− -double-knockout mice in the regeneration-phase despite reduced liver damage in histological and biochemical analysis. Currently, the molecular mechanism of liver regeneration by c-myc is investigated. Conclusions: C-myc does not affect liver injury in Fah−/− -mice during severe and moderate liver-damage. Surprisingly, loss of c-myc ameliorates liver injury and enhances liver regeneration during repeated flares of liver injury. Long-term studies will reveal c-myc’s contribution to hepatocarcinogenesis in the Fah-model. 418 SEPSIS INDUCED DAMAGE IN THE LIVER IS MODULATED BY PEROXIREDOXINS N. Huber1 , T. Annecke2 , A. Hillenbrand1 , D. Henne-Bruns1 , U. Knippschild1 , J. Tschop ¨ 2 . 1 General-, Visceral- and Transplantation Surgery, University of Ulm, Ulm, 2 Anesthesiology, University of Munich, Munich, Germany E-mail: [email protected] Background and Aims: Severe sepsis is characterized by hyperinflammation, oxidative damage, hypercoagulation, tissue hypoperfusion, hypoxia, and immune suppression, leading to multiorgan dysfunction. During sepsis, liver dysfunction is common and contributes to high mortality. Although the precise mechanisms by which the liver is affected are unclear, evidence is increasing that failure of mitochondria to effectively couple oxygen consumption with energy production contribute to the pathology of sepsis. An imbalance between formation and clearance of reactive oxygen species (ROS) results in oxidative stress and significantly contributes to dysfunction of cells during sepsis. In addition, ROS are essential to various functions of cells. However, moderate levels of antioxidant defenses are required to protect against the harmful effects of excessive ROS production. The aim of this study was to evaluate the levels of antioxidants, especially those of peroxiredoxin (Pdrx) isoforms 3 and 6, during induced sepsis in a mouse model. Material and Methods: For all experiments 8–12 weeks old male C57BL/6 mice either on a normal or high fat diet were used. Mice were randomized for the groups and all surgical procedures were performed in a matched experimental setting. Sepsis was induced by cecal ligation and puncture method. Animals were sacrificed 6 h, 12 h, and 24 h after induction of sepsis. Changes in the expression of various proteins were analyzed by Western Blot analyses and immunhistochemistry. Additionally, Kaplan-Meieranalysis were performed. Results: Already 6 h upon induction of sepsis changes in the cytoplasm of the liver could be detected. The expression levels of the relatively new group of antioxidants, the peroxiredoxins (Prdx 3 and 6) in the liver were detected by Western blot analyses and immunohistochemistry. Furthermore, we show that the influence of nutrition modulate the levels of Prdx proteins in the liver thereby influencing the severity of sepsis and survival. In addition, our results show that changes in the expression of Prdx isoforms modulate death signalling. Conclusions: Here we show for the first time that changes of Pdrx levels in the liver depend on the severity of sepsis. These results might contribute to the development of new therapeutical strategies in the treatment of severe sepsis.
419 RESULTS OF AUTOLOGOUS BONE MARROW CELL INFUSION IN PATIENTS WITH ADVANCED LIVER CIRRHOSIS J.K. Kim1,2 , H.J. Chung1 , D.H. Jung1 , D.Y. Kim1,2 , S.H. Ahn1,2 , C.Y. Chon1 , K.-H. Han1,2 , K.S. Lee1,2 . 1 Dept. of Internal Medicine, Yonsei University College of Medicine, 2 Liver Cirrhosis Clinical Research Center, Seoul, Republic of Korea E-mail: [email protected] Background and Aim: Liver cirrhosis (LC) is the end stage of chronic liver disease and is very difficult to treat. Although human clinical trials of bone marrow cell (BMC) infusion for cirrhotic patients showed positive results, the long-term follow up data after the procedure is limited. The aim of this study is to analyze the clinical results of phase II study after autologous BMC infusion (ABMI) in patients with advanced LC. Patients and Methods: Patients aged between 18 and 75 with a clinical diagnosis of advanced LC (Child-Pugh class B) and no viable hepatocellular carcinoma on magnetic resonance imaging (MRI) underwent ABMI. Autologous BMCs were harvested from the ilium under general anesthesia, and infused into peripheral vein after RBC depletion and mononuclear cell concentration. Serologic tests and questionnaires about performance were repeated during the 12-month period after the ABMI. Data after liver transplantation were not included. Results: Twenty patients were screened and 19 patients were enrolled. Mean age was 52 year-old. Patients (M:F = 9:10) were followed for more than 12 months after ABMI. Eighteen patients had B-viral liver cirrhosis and one had alcoholic liver cirrhosis. Serum albumin level was improved from 2.9 (2.3–3.6) g/dL before ABMI to 3.4 (2.6–4.5) g/dL at 12th month after ABMI. Prothrombin time showed improving tendency from 70 (60.0–84.0) % before ABMI to 77 (69.0–100) % at 12th month. Hemoglobin was increased from 11.6 (9.5–12.5) g/dL before ABMI to 13.25 (11.4–16.8) g/dL at 12th month. Child-Pugh score was decreased from 7.6 (7–12) before ABMI to 6.8 (5–10) at 12th month after ABMI. Performance status score improved from 78.9 (70–80) before ABMI to 90.8 (70–100) at 12th month. Conclusions: ABMI for advanced liver cirrhosis improved hepatic function and ABMI can be a bridge therapy waiting liver transplantation. 420 NON-PARENCHYMAL CELLS REGULATE PROLIFERATION AND DIFFERENTIATION OF HUMAN LIVER PROGENITOR CELLS J.W.C. Kung, I.S. Currie, P.W. Hadoke, S.J. Forbes, J.A. Ross. University of Edinburgh, Edinburgh, UK E-mail: [email protected] Background: Human foetal liver progenitor cells (LPCs) have promise for liver cell therapy, however, culture of LPCs alone result in inadequate functionality and phenotypic stability. The conditions required for LPC proliferation and differentiation are currently unknown. We hypothesised that non-parenchymal cells (NPCs) regulate proliferation and differentiation of LPCs. Methods: EpCAM+ LPCs were isolated by immunoselection from second trimester human livers. NPCs were isolated through EpCAM negative selection. LPCs, NPCs and mixed LPC/NPC cultures were grown in Williams’ Medium E (WME) and NPC-conditioned medium (NPC-CM) for 13 days and their hepatocytic functions were compared. Results: EpCAM+ cells had hepatoblast phenotype (AFP/Albumin/ CK19 positive). EpCAM− NPCs consisted of tube forming endothelial cells and mesenchymal cells. Mixed LPC/NPC cultures had significantly greater synthetic competence than LPCs alone (twoway ANOVA, p < 0.01). We confirmed this was mediated through soluble factor(s) by performing media transfers. LPCs grown in NPC-CM had greater hepatocytic maturity by secreting significantly
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